ISOLATION OF ARCANOBACTERIUM HAEMOLYTICUM FROM
PATIENTS WITH PHARINGITIS
, Micaela Scripcaru1
, Angela Romaniuc1
, Mariana Dumbrava1
, Paraschiva Onu2
1. Institute of Public Health Iasi, 2. Health Centre “Nicolina” Iasi
Abstract. Arcanobacterium haemolyticum has been described as a rare etiologic agent in acute pharingotonsilitis in pediatric and young population.Four strains of A. haemolyticum were isolated from throat swabs of 3584 patients (10-26 years ) with acute pharingitis. The samples, collected in January 1997-January 2001, were plated on Mueller-Hinton agar with 10 % sheep blood and incubated 24-48 hours at 37 °C.
In three cases A.haemolyticum was the only one etiologic bacterial agent and in the forth case association of A.haemolyticum with S.aureus was present.
The identification of A.haemolyticum strains was based on cells and colonies’ morphology, cultural and biochemical characters.
Key-words: Arcanobacterium haemolyticum, isolation, pharingitis.
Rezumat. A.haemolyticum a fost semnalat, cu frecvenţă redusă, ca agent etiologic al faringoamigdalitelor acute la copii şi tineri. Autorii descriu 4 tulpini de A.haemolyticum izolate din exudate faringiene de la bolnavi cu faringoamigdalită acută .3484 probe, recoltate in perioada ianuarie 1997-ianuarie 2001,au fost insămânţate pe agar Mueller- Hinton suplimentat cu 10 % sânge de berbec şi incubate la 37 °C timp de 24– 48 ore.
La 3 din cazuri A.haemolyticum a fost unicul agent izolat iar la 1 caz a fost prezent în asociaţie cu S.aureus.
Tulpinile de A.haemolyticum au fost confirmate pe baza morfologiei celulelor bacteriene şi a caracteristicilor culturale şi biochimice.
Cuvinte – cheie : Arcanobacterium haemolyticum, izolare, faringită.
haemoliticum) is an aerobic gram –
positive rod that was first described by
MacLean et al., in 1946, isolated in
persons with pharingitis and skin
infections. Recently, many studies
point out that A.haemolyticum has
been isolated from patients with
pharingitis and various other infections in
Europe, USA and Asia (1,2 ).
The taxonomical position of
A.haemolyticum caused many confusions
because it is a close phenotypical similarity
to Actinomyces pyogenes (C. pyogenes ), an
animal bacterial pathogen (1). From
1982 this species was classified as A.
haemolyticum by Collins et al., in a
new genus Arcanobacterium composed
of this single species (1,3, 4).
In the infection with A.haemolyticum
clinical most cases involve pharingitis
and / or tonsilitis and approximately
50% are exudative. Throat infections
are often accompanied by cervical
lymphadenopathy (1,2,3,5). Simptoms
resemble those of beta – haemolytic
streptococci or viral infection.
An erythematous morbilliform or
scarlatinal rush of trunk, neck or
extremities were associated with the
presence of A. haemolyticum. In addition,
central nervous system infections, sepsis,
endocarditis, osteomyelitis, dermatologic
and other infections were described
The aim of this study were to determine
the presence of A.haemolyticum in acute
pharingitis in children and young
population and to characterize the
A.haemolyticum isolated strains.
MATERIAL AND METHODS
3584 throat culture samples were
collected during January 1997–
January 2001 from patients with acute
pharingitis. The patients of 5–26 years
of age were seen in districtual or
Children and Students Hospital of Iasi
The samples were plated on Mueller –
Hinton agar added with 10% sheep
blood and incubated for 24–48 hours
at 37 C in aerobic conditions.
The A. haemolyticum strains were
identified by the following tests : the
small colonies (< 1 mm diameter) with
incomplete beta–haemolysis on sheep
blood agar, Gram stain, catalase,
nitrate reduction, urease, gelatin
hydrolysis, reverse CAMP, DNase
test, fermentation of glucose, lactose,
sucrose, maltose, mannitol and xylose
– table 1. The tests Voges–Proskauer
diffusion method. They were tested to
penicillin, erythromycin, tetracycline,
gentamycin, cephalotin, ceftazidime,
ceftriaxone, chloramphenicol, and
RESULTS AND DISSCUSION
A.haemolyticum was isolated in 4
(0.1%) cases of all investigated
patients (10–26 years). In 3 cases this
species was the only bacterial
pathogen and in one case was
associated with S.aureus .
Clinical manifestations of A.haemolyticum
pharyngitis were similarly with those of
streptococcal pharyngitis but the severity
of the disease varied.Sore throat and
pharyngeal erythema were always
present. Additional symptoms and signs
included fever, non–productive cough and
headache. Another symptoms and signs
such as skin rash, tonsillar exudates,
lymohadenopathy have also been
described. Erythema multiforme and
urticarial rash have also been found in
patients with A.haemolyticum pharyngitis
(1,6,7). In our investigation the frequency
of A. haemolyticum isolation was very
low. The isolation rate of this
pathogen from clinical throat
specimens varies from 0.07% to 1.3%
in non-age selected material (1). In
Romania were described 24 cases
confirmed with A.haemolyticum (6,7).
Epidemiological data suggest that
A.haemolyticum pharyngitis is
primarly a disease of adolescent and
young adults. Carlson P.; Coman et
on classical culture techniques.This
organism grows slow, with little (<1
mm diameter) and smooth/rough
colonies on blood agar and it is easly
overlooked in the bacteriological
laboratory. Around of the colonies
appears a narrow incomplete haemolysis
zone. Banck et al., considered that the
detection of beta- haemolysis
produced by this bacteria can be
facilitated by using special
double-layered human blood agar plates (3 ).
The identification of A.haemolyticum
was made by classical biochemical
tests presented in table 1 and 2. The
catalase and gelatin hydrolysis
differentiated this species of
Corynebacterium sp. and Actinomyces
pyogenes. A catalase negative,
gram-positive rods which is reverse CAMP
and DNase positive tests is very
important for the identification of
A.haemolyticum. The inclusion of
bacteria in genus and species have
been confirmed by the acid production
test from maltose, lactose, sucrose,
xylose and mannitol .
Many authors succesfully used the
API Staph and API Coryne systems
for testing carbohydrate fermentation
by A.haemolyticum (1,6,8).
This species can be divided in smooth
and rough biotypes by biochemical
tests. The majority smooth biotype
strains fermented sucrose and/or
trehalose in API Staph but not
produced beta-glucoronidase. The
rough biotype strains produced
beta-glucoronidase but not fermented
sucrose and trehalose.
All our A. haemolyticum strains were
susceptible to penicillin, erythromycin,
cephtazidime,cephtriaxone.2 strains were
resistant to tetracycline and the
no activity in vitro. In majority
studies, the A. haemolyticum strains
were susceptible to the antibacterial
agents recommended for treatment of
streptococcal tonsillitis, such as penicillin,
oral cephalosporins, erythromycin and
clindamycin (1,3,6, 8).
y A.haemolyticum was isolated from
0.1% of the throat specimens of
10-26 year old patients with acute
pharingitis, using sheep blood agar
on which this species grows as little
smooth or rough colonies.
y The identification of this organism
was possible using: Gram stain,
catalase, reverse CAMP, gelatin
hydrolysis and the capacity of acid
formation from glucose, maltose,
sucrose, xylose, mannitol .
y All A.haemolyticum strains were
susceptible to penicillin, cephalosporins,
macrolides, tetracycline and resistant to
Table 1. Differential characteristics of A. haemolyticum and other bacterial groups Acid from Bacterial group Catal ase M ot ili ty Nitrat reducti on Ureas e Gel ati n hy dr olys is C arb ohyd ra te ferm ent ati on Gl ucose Esculin hydr olyz ed Mal tos e L acto se Su cr os e Xyl os e Mann itol Corynebacterium sp. + - V V V + + - V - V - - Arcanobacterium haemolyticum (C. haemolyticum) - - - - - + + - + + V - - Actinomyces pyogenes (C. pyogenes) - - - - + + + - V + V + V Listeria monocytogenes + + - - - + + + + V V - -
+, positive; -, negative; V, variable reaction.
Table 2. Principal characteristics of A. haemolyticum
Test Reaction Test Reaction β-haemolysis Catalase Nitrate reduction Urease Gelatin hydrolysis Esculin hydrolysis + - - - - - Reverso CAMP DNase Glucose Maltose Sucrose Xylose + + + + V -
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